Custom RT2 Profiler PCR Arrays are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

Positive results with as little as 25 ng RNA.

Different amounts of universal total RNA were characterized using the Human Inflammatory Cytokines and Receptors RT2 Profiler PCR Array and the percentage of detectable genes calculated for each amount. As little as 25 ng total RNA yields an >80% positive call, even for cytokines expressed at very low levels.

Simple procedure.

A single gene-specific product in every reaction.

Universal total RNA was characterized using the TGFβ/BMP Signaling Pathway RT2 Profiler PCR Array, followed by dissociation (melting) curve analysis. The PCR array specifically detects individual genes, despite the expression of related gene-family members in the same RNA sample.

Available plate layouts.

PCR arrays yield highly accurate results.

A representative set of assays for 4000 genes used in RT2 Profiler PCR Arrays have an average amplification efficiency of 99% with a 95% CI from 90–110%. Consistently high amplification efficiencies enable PCR arrays to accurately analyze multiple genes simultaneously utilizing the ΔΔCT method.

High reproducibility among different users.

4 replicate sets of raw threshold data (1–4) obtained by 2 different scientists (A and B) at 2 different times using Human Drug Metabolism RT2 Profiler PCR Arrays were directly compared. The results demonstrate a high degree of correlation (R2 >0.990).

Performance

Sensitivity

With the sensitivity of the RT2 First Strand Kit, as little as 1 ng or as much as 5 µg of total RNA per array plate provides greater than 80% present call rates (see figure "Positive results with as little as 25 ng RNA").

Reproducibility

The complete PCR array system demonstrates strong correlations across technical replicates, lots and instruments with average correlation coefficients >0.99, ensuring reliable detection of differences in expression between biological samples (see figure "High reproducibility among different users").

Specificity

The PCR array system, with high-quality input RNA, is guaranteed to yield single bands of the predicted size without primer-dimers or other secondary products, therefore providing highly accurate real-time PCR results (see figure "A single gene-specific product in every reaction").

Uniform PCR amplification efficiency

Uniform PCR amplification efficiency is required for PCR array technology to allow accurate comparison of gene expression across all genes. The unique combination of a proprietary primer design algorithm and rigorous testing of every primer assay guarantees the high performance of every primer assay on PCR arrays (see figure "PCR arrays yield highly accurate results").

This combination provides Custom RT2 Profiler PCR Arrays with the specificity and the high amplification efficiencies required for accurate real-time SYBR Green results. PCR arrays are easy to use in any research laboratory.

Available formats

Custom RT2 Profiler PCR Arrays are available in the following plate layouts (see the tables below and figure "Available plate layouts". Select the genes and the plate format required.

Rotor-Gene and 96-well plate or 100-well disc custom qPCR arrays

Format

Number of plates (minimum)

8 genes, 12 samples per plate

12

12 genes, 8 samples per plate

12

16 genes, 6 samples per plate

12

24 genes, 4 samples per plate

24

32 genes, 3 samples per plate

24

48 genes, 2 samples per plate

24

96 genes, 1 sample per plate

24

All formats

Per additional 12 array

384-plate custom qPCR arrays

Format

Number of plates (minimum)

8 genes, 48 samples per plate

6

12 genes, 32 samples per plate

6

24 genes, 16 samples per plate

6

32 genes, 12 samples per plate

6

48 genes, 8 samples per plate

6

64 genes, 6 samples per plate

6

96 genes, 4 samples per plate

6

128 genes, 3 samples per plate

6

192 genes, 2 samples per plate

24

384 genes, 1 sample per plate

24

All formats

Per additional 6 array

Arrays are available in additional formats or with special assay designs (position, variants, etc.), please inquire.

Procedure

Simply mix the cDNA template with the appropriate ready-to-use PCR master mix, aliquot equal volumes to each well of the same plate, and then run the real-time PCR cycling program (see flowchart "Simple procedure"). Custom RT2 Profiler PCR Arrays are compatible with all QIAGEN, ABI, Bio-Rad, Eppendorf, Roche and Stratagene instruments.

Flexible layout and controls

Custom RT2 Profiler PCR Arrays are available in 96-well plate, 384-well plate and 100-well disc formats, and are used to monitor the expression of up to 84 or 370 genes related to a specific research area of interest, plus 5 housekeeping genes. Custom RT2 Profiler PCR Arrays may also include control elements for:

Data normalization

Genomic DNA contamination detection

RNA sample quality

General PCR performance

Easy-to-use data analysis

Data can be analyzed using an easy-to-use Excel-based data analysis template or web-based software. Data analysis is based on the ΔΔCT method with normalization of the raw data to either housekeeping genes.

Applications

Custom or Modified RT2 PCR Profiler Arrays offer a robust platform for biomarker screening and validation and can be used in all areas of biological and medical research, including: